Biomedical subjects

D C Straus

Publications and source records attributed to D C Straus.

At least 55 records · Page 3Linked to original sources

PubMed1990

Immune response to pulmonary injection of Pasteurella haemolytica-impregnated agar beads followed by transthoracic challenge exposure in goats.

A method of inducing Pasteurella haemolytica serotype 1 (Ph1) lung infection in goats, using low numbers of bacteria and without impairing host immunity, was developed. Two trials were conducted. Results of trial 1, using 10 principals (Ph1 agar beads) and 6 controls (agar beads alone), indicated that Ph1 organisms imbedded in agar beads could survive host lung defenses for 32 days. Results of trial 2 indicated that lung immunity in the inoculated goats (principals) was high and they were more protected than controls against a transthoracic challenge of Ph1 (1.18 x 10(7) colony-forming units) injected into a lung of each goat on posttreatment day 35. When comparing challenge-exposed principals with controls, the controls developed rectal temperatures above normal for a longer time, duration of anorexia was longer, and signs of depression were seen. The controls developed large areas of consolidated lung tissue, more Ph1 isolates were recovered from nasal turbinates and lung tissue, and higher Ph1 concentrations were found in the lungs. The serum Ph1 indirect hemagglutination antibody titers in the principals of both trials increased, compared with titers in controls. Principal goats in trial 2 had higher Ph1 indirect hemagglutination antibody titers after injection of Ph1-impregnated agar beads and less severe lung lesions after challenge exposure than did controls. The small pneumonic consolidated lesions in the principals, compared with extensive lesions in controls after Ph1 challenge exposure, indicated a high degree of immunity after exposure to Ph1 organisms imbedded in agar beads.

Animals↗

PubMed1989

Production of an extracellular toxic complex by various strains of Pseudomonas cepacia.

Six isolates of Pseudomonas cepacia, representing various serotypes of the organism and possessing similar degrees of virulence in mice, were examined for their production of an extracellular toxic complex (ETC) in vitro. This compound is lethal for mice and produces extensive lung pathology in rats; it is composed of a surface carbohydrate antigen, lipopolysaccharide and protein. All six isolates produced the ETC. The LD50 values for the six ETC preparations ranged from 395 micrograms for strain 61g to 1750 micrograms for strain 90ee. Only two of the six ETC preparations contained ketodeoxyoctonate detectable by the methods used, and these two were the most toxic. Rabbit antiserum to the ETC of a serotype D strain could significantly protect mice only against serotype D strains. Examination of the various phases of growth of P. cepacia showed that there was extracellular release of the ETC beginning in the early logarithmic phase and continuing through the late stationary phase. The presence of the ETC in the supernatant fluids was due to release of this material rather than to cell lysis. In addition, at least one strain of P. cepacia was shown to produce an alginic acid-like compound.

Animals↗

PubMed1988

Bactericidal activity of testicular macrophages.

The purpose of these studies was to determine if testicular macrophages are capable of bactericidal activity. Testicular macrophages were isolated from adult Wistar rats and studied in vitro. Studies were designed to determine if these cells could kill pathogenic gram-negative organisms and if these cells secreted lysozyme, an enzyme involved with the lysis of the cell wall of gram-positive bacteria. The regulation of lysozyme secretion by hormones and lipopolysaccharide was also studied. The secretion of this enzyme by testicular macrophages was also compared to enzyme secretion by macrophages isolated from other tissues. We also studied the secretion of superoxide anion, which is known to be involved in cytotoxic reactions. It was found that testicular macrophages were capable of killing up to approximately 38% of a virulent encapsulated strain of Klebsiella pneumoniae within 1 h. This process was in part dependent upon the presence of immune serum generated against these organisms but could not be mimicked by control serum or immune serum tested in the absence of macrophages. Testicular macrophages secreted lysozyme in culture for at least 8 days; however, macrophages from the peritoneal cavity and lung secreted significantly more lysozyme under the same conditions. Lipopolysaccharide suppressed lysozyme secretion in a dose-dependent manner, whereas neither follicle-stimulating hormone, testosterone, nor leuteininzing hormone had an effect on lysozyme secretion. Finally, testicular macrophages secreted superoxide anion in a manner similar to peritoneal macrophages. These studies indicate that testicular macrophages have the capability to mount an appropriate defense against pathogenic bacteria by opsonization-dependent phagocytosis, the secretion of lysozyme, and the production of super oxide anion.

Animals↗

PubMed1988

The importance of extracellular antigens in Pseudomonas cepacia infections.

A clinical isolate of Pseudomonas cepacia from a cystic fibrosis patient was examined for its ability to produce extracellular toxic material. The organism was grown to stationary phase in a defined medium and toxic material was isolated by ultrafiltration, ion-exchange chromatography on DEAE-Sephacel and gel-filtration chromatography on Sepharose 4B. It consisted of a surface carbohydrate antigen, lipopolysaccharide and protein, and had an LD50 (when injected intraperitoneally into mice) of 395 +/- 20 micrograms. The toxicity appeared to be associated with the lipopolysaccharide portion of the complex, because boiling for 15 min and exposure to proteolytic enzymes had no effect on toxicity. However, saponification destroyed the toxicity of the compound. Studies employing radial immunodiffusion with the sera of mice infected with this organism demonstrated production of the complex in vivo at levels approaching those sufficient to produce death. When sublethal amounts of this complex were placed in the lungs of specific-pathogen-free rats, the lung pathology observed after 12, 24, 36 and 48 h was extensive. However, antibody generated in rabbits against this material could protect mice against the complex, as well as against challenge by the homologous organism. These data indicate that extracellular toxic material produced by P. cepacia may be responsible for the lethality and lung tissue destruction normally associated with an active pneumonia caused by this organism.

Animals↗

PubMed1988

Production of lipase by clinical isolates of Pseudomonas cepacia.

Ten clinical isolates of Pseudomonas cepacia from the sputum of cystic fibrosis patients were examined for the ability to produce lipase. Lipase substrates used included egg yolk agar, four different polyoxyethylene sorbitans (Tweens), and p-nitrophenylphosphorylcholine, a chromogenic substrate used to assay for phospholipase C. Lipase activity was detected in the filtrates of organisms grown to the exponential phase in either tryptose minimal medium or chemically defined medium. Lipase activity increased in the filtrates if the cultures were allowed to proceed into the stationary phase. None of the isolates produced phospholipase C. Lipase activity on Tween 20 ranged from 41.6 X 10(-3) to 640.0 X 10(-3) U/micrograms of protein. The activity was similar or slightly lower when Tween 40, 60, or 80 was used as the substrate. There was no correlation between lipase activity on Tween and that demonstrated on egg yolk agar. Lipase activity increased as pH increased from 7.0 to 9.0. Boiling for 5 min resulted in 66% loss of enzyme activity. The remaining activity continued to decrease with increasing boiling time. The enzyme was purified by gel filtration on Sephadex G-200, and the resultant preparation, when subjected to polyacrylamide gel electrophoresis, resulted in a single protein band (molecular weight, approximately 25,000) from which lipase activity could be eluted. The purified lipase was not cytotoxic to HeLa cells, nor was it toxic when injected intravenously into mice.

Animals↗

PubMed1987

Characterization of neuraminidases produced by various serotypes of group B streptococci.

Neuraminidase produced by 11 strains of group B streptococci (GBS), from serotypes Ia, Ib, Ic, II, and III, were characterized according to molecular weight, antigenic identity, and substrate specificity. Following growth in a chemically defined medium, ammonium sulfate-concentrated culture supernatants were assayed for activity with bovine submaxillary mucin as substrate. Neuraminidase produced by GBS strain 122 (serotype III) was purified by a combination of salt fractionation, affinity chromatography with Affi-Gel Blue, ion-exchange chromatography with DEAE-cellulose, and gel filtration on Sephadex G-200. Purified neuraminidase was used to immunize rabbits, and the resultant antiserum reduced the activity of purified neuraminidase from strain 122 by 87.7%. The antiserum also reduced the activity of neuraminidases produced by the other four serotypes by between 78.3 and 90%. Molecular weight estimates of the neuraminidases produced by the various serotypes were obtained by gel filtration chromatography on Sephadex G-200. The molecular weights obtained for the neuraminidases from the representative strains of each serotype ranged from 110,000 to 180,000. In addition, all of the GBS neuraminidases examined (regardless of the producing serotype) were active only on bovine submaxillary mucin. On the basis of these results, it appears that the neuraminidases produced by different GBS serotypes are quite similar.

Antibodies, Bacterial↗

PubMed1987

Production of an extracellular toxic complex by various strains of Klebsiella pneumoniae.

Six isolates of Klebsiella pneumoniae (two serotype 1 isolates and a capsular variant of one of these, and two serotype 2 isolates and a capsular variant of one of these) possessing various degrees of virulence in rats and mice were examined for their in vitro production of an extracellular toxic complex (ETC). The ETC has been shown to be lethal for and produce extensive lung pathology in mice. This compound has been shown to be composed of capsular polysaccharide, lipopolysaccharide, and a small amount of protein. All six isolates produced the ETC. Immunization of experimental animals with sublethal doses of the ETC was protective against both homologous and heterologous strains, and this protection was due to antibody production. An examination of the various phases of growth of K. pneumoniae showed that there was extracellular release of the component parts of the ETC occurring during all phases of growth. The presence of the ETC in the supernatant fluids was due to actual release of this material as opposed to cell lysis. Antibodies to the lipopolysaccharide portion (which has been shown to possess the observed toxicity) of the ETC were protective against the homologous bacterium.

Animals↗

PubMed1986

Biochemical and immunological characterization of the variant surface coat glycoprotein shed by African trypanosomes.

As the variant surface coat glycoprotein (VSG) was shed from Trypanosoma brucei rhodesiense into the blood of infected rats, it was biochemically characterized and compared with VSG that had been purified from trypanosomal homogenates. To determine if VSG was in association with lipid, membranes and lipoproteins in plasma of infected rats (IRP), VSG isolated from plasma (PVSG), and VSG isolated from trypanosomal homogenates (HVSG) were all concentrated by ultracentrifugation and assayed for the presence of VSG by radial immunodiffusion (minimum level of detection, 25 micrograms/ml) and by immunoelectroblots (minimum level of detection, 1 microgram/ml). Crimson red was used to detect lipid (minimum level of detection, 10 micrograms per sample) in electrophoresed samples. The VSG was neither concentrated with membrane or lipoprotein fractions nor stained by lipid crimson. Lipids from normal rat plasma, IRP, trypanosomal homogenates, HVSG, and PVSG were also extracted and separated by thin-layer chromatography (minimum level of detection, 20 micrograms of trypanosomal phospholipid per sample). The trypanosomal homogenates had five bands as detected by iodine vapors, of which three were phospholipids as detected by molybdenum blue. Both normal rat plasma and IRP had identical patterns of bands with a single phospholipid. The PVSG had one neutral lipid contaminant that apparently was not physically associated with the shed surface coat. The HVSG contained no lipids at all. Therefore, no evidence was obtained to implicate an association between membranes and VSG, once the latter had been shed into the blood of infected hosts. From immunoelectroblots of denatured material, it was determined that both HVSG and PVSG had the same reduced molecular weight. From molecular sieve column chromatography, however, it was determined that VSG released during the homogenization of trypanosomes is a noncovalently linked dimer, whereas that shed in the blood is apparently a trimer. This difference in native structure made no difference in immunological effect. Administered in a regimen that mimicked what the host encounters during a first peak of parasitemia, both HVSG and PVSG induced nonspecific proliferation of splenic lymphocytes and production of unelicited antibodies without the generation of nonspecific immunosuppression. This polyclonal activation of lymphocytes was not the result of contamination by exogenous pyrogen, because the activity was lost if VSG was immunologically absorbed from plasma.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

PubMed1985

Importance of a lipopolysaccharide-containing extracellular toxic complex in infections produced by Klebsiella pneumoniae.

A Klebsiella pneumoniae serotype 2 strain was examined for its ability to produce extracellular toxic material. The organism was grown to the stationary phase in a defined medium, and the toxic material was isolated by ultrafiltration-ion-exchange chromatography on DEAE-Sephacel and gel filtration chromatography on Sepharose 4B or 2B. It was found to be comprised of 63% capsular polysaccharide, 30% lipopolysaccharide, and 7% protein and possessed a 50% lethal dose (when injected intraperitoneally into mice) of 393 +/- 45 micrograms. The toxicity appeared to be associated with the endotoxin portion of the compound, because boiling for 15 min and exposure to proteolytic enzymes had no effect on the toxicity. However, saponification destroyed the toxicity of the compound. Studies employing radial immunodiffusion examining the sera of mice infected with this organism demonstrated in vivo production of the complex at levels sufficiently high to produce death. When sublethal amounts of this complex were placed in the lungs of specific-pathogen-free mice, the lung pathology observed after 24, 48, and 72 h was similar to the damage caused by an active K. pneumoniae lobar pneumonia. These data indicate that this extracellular toxic compound produced by K. pneumoniae may be responsible for the lethality and lung tissue destruction normally associated with an active lobar pneumonia caused by this organism.

Animals↗

PubMed1985

Extracellular polysaccharide production by Klebsiella pneumoniae and its relationship to virulence.

Klebsiella pneumoniae serotype 1 and serotype 2 and their capsular variants were examined for production of cell-associated capsular polysaccharides and extracellular capsular polysaccharides. The virulence of these organisms in experimental animals was examined via intraperitoneal injection in mice and transtracheal inoculation into the lungs of rats. It was found that the production of either polysaccharide component correlated with the observed virulence. The extracellular polysaccharides were purified by ethanol precipitation, electrodialysis, extraction with quaternary ammonium salts, and gel filtration. These purification steps allowed for the separation and purification of both the extracellular lipopolysaccharide and the extracellular capsular polysaccharide. Purified extracellular capsular polysaccharide and extracellular lipopolysaccharide were co-injected with K. pneumoniae intraperitoneally into mice to determine if either of these substances would produce an effect on the natural course of infection in these animals. These studies showed that only purified extracellular lipopolysaccharide enhanced the virulence of K. pneumoniae when co-injected into mice, and this virulence enhancement correlated with the content of extracellular lipopolysaccharide, but not extracellular capsular polysaccharide in mixtures of these polysaccharides. Saponification of K. pneumoniae serotype 1 extracellular polysaccharides significantly decreased their virulence-enhancing capabilities in mice, further suggesting that extracellular lipopolysaccharide may play a role in these infections.

Animals↗

PubMed1983

Neuraminidase production by a Streptococcus sanguis strain associated with subacute bacterial endocarditis.

The properties of an extracellular neuraminidase produced by a Streptococcus sanguis strain (isolated from a confirmed case of subacute bacterial endocarditis) during growth in a defined medium was examined in this investigation. This enzyme, isolated from concentrated culture supernatants of S. sanguis biotype II, was active against human alpha-1 acid glycoprotein, N-acetylneuramin lactose, bovine submaxillary mucin, and fetuin. Neuraminidase production paralleled bacterial growth in defined medium and was maximal in the early stationary phase of growth but decreased dramatically, probably owing to protease production, during the late stationary phase. The enzyme was purified to near homogeneity by a combination of salt fractionation, ion-exchanged chromatography on DEAE-Sephacel, and gel filtration on Sephadex G-200. These procedures yielded an enzyme preparation that possessed a specific activity of 174.4 mumol of sialic acid released per min per mg of protein against human alpha-1 acid glycoprotein. The Km value for this enzyme with human alpha-1 acid glycoprotein as substrate was 2.5 X 10(-3) M, and the enzyme possessed a pH optimum of 6.5. The S. sanguis neuraminidase had a molecular weight of approximately 85,000 as estimated by gel filtration and approximately 90,000 when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme was stable at temperatures of 4 and 37 degrees C for 3 h, but approximately 50% of the enzymatic activity was lost within 30 min at 50 degrees C, with 100% of the enzymatic activity being destroyed within 10 min at temperatures of greater than or equal to 65 degrees C.

Endocarditis, Subacute Bacterial↗

PubMed1983

Pseudomonas aeruginosa exoproducts as pulmonary virulence factors.

An experimental animal model of chronic pseudomonas pneumonia was used to document the production of potential virulence factors by Pseudomonas aeruginosa during the infection. The production of exotoxin A, proteolytic enzymes, and the serotype-specific lipopolysaccharide and slime-layer antigens during the infection was examined by solid-phase radioimmunoassay of serum from infected rats and by indirect immunofluorescence tests of their lung tissue. Rats inoculated intratracheally with purified bacterial exoproducts, delivered alone or in combination, developed pulmonary histopathology similar to that induced by the experimental infection. The results indicate that these exoproducts are produced during the course of the pulmonary infection and suggest that they are involved in the observed lung pathology.

ADP Ribose Transferases↗

PubMed1982

Immunogenic fractions of Cryptococcus neoformans.

Cryptococcus neoformans cell and culture supernatant extracts were fractionated by ion exchange and gel filtration column chromatography. Various fractions were used to immunize mice, and to assess the release of migration inhibition factor, delayed type hypersensitivity, and protective immunity after challenge with C. neoformans. Results suggest that the C. neoformans fractions, which protect mice, contain a high molecular weight, predominantly carbohydrate antigen that can be distinguished from the capsular polysaccharide.

Animals↗

PubMed1982

Lobar pneumonia in rats produced by clinical isolates of Klebsiella pneumoniae.

Transtracheal instillation of clinical isolate Klebsiella pneumoniae serotype 1 (KP1) into the lungs of rats resulted in the production of a characteristic, chronic lobar pneumonia. To further examine this phenomenon, two variants of this organism were employed in this experimental model. These variants differed markedly in capsule size, colony morphology, and in virulence, as determined by mouse lethality tests. The ability of these strains to establish a lobar pneumonia in rats correlated with the virulence of the respective organisms as monitored by intraperitoneal injection in mice. The 50% lethal doses in mice were 4.9 x 10(1) colony-forming units (CFU) for the more virulent KP1 strain (KP1-O) and 1.42 x 10(5) CFU for the less virulent variant (KP1-T). In the rat lung model, marked lung pathology was evident by day 6 with a KP1-O inoculum of 5 x 10(2) CFU, whereas KP1-T caused little or no lung pathology when delivered transtracheally at a concentration of 7 x 10(6) CFU. Two relatively nonvirulent variants of K. pneumoniae serotype 2 were also used in this rat lung model and were found not to produce a lobar pneumonia even when delivered in large doses. These results indicate that a chronic lobar pneumonia can be established in a rat model if the appropriate organism is employed and the virulence of K. pneumoniae injected intraperitoneally into mice is an excellent indicator of an organism's potential to cause lobar pneumonia in rats.

Animals↗

PubMed1982

Protease production by Streptococcus sanguis associated with subacute bacterial endocarditis.

A viridans streptococcus (Streptococcus sanguis biotype II) isolated from the blood of a patient with subacute bacterial endocarditis was examined for protease production. In broth culture, extracellular proteolytic enzymes were not produced by this organism until after the early exponential phase of growth, with maximal protease production occurring during the stationary phase. Four distinct proteases were isolated and purified from the supernatant fluids of stationary-phase cultures, employing a combination of ion-exchange column chromatography, gel filtration column chromatography, and polyacrylamide gel electrophoresis. All four proteases could be eluted from a diethylaminoethyl cellulose column at a sodium chloride gradient concentration of 0.25 M but were separable by gel filtration chromatography on a Sephadex G-100 column. They varied in molecular weights as determined by gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis from approximately 13,000 to 230,000. All four proteases had pH optima of between 8.0 and 9.0, and two of the proteases were active against casein, human serum albumin, and gelatin but were not active against elastin and collagen. The remaining two proteases were able to degrade only casein and gelatin. These results show that S. sanguis is able to excrete maximal levels of potentially destructive enzymes when the organisms are not actively multiplying. This finding may explain some of the damage caused in heart tissue by these organisms during subacute bacterial endocarditis.

Caseins↗

PubMed1982

Antibody response to Pseudomonas aeruginosa exoproducts in cancer patients.

We analyzed sera from 28 patients with various types of malignancies for the occurrence of antibodies against exotoxin A of Pseudomonas aeruginosa and two Pseudomonas proteases. A total of 27 of these individuals were colonized or infected with P. aeruginosa at one time or another during the study, whereas the remaining patient was colonized with four non-P. aeruginosa species of Pseudomonas. Sera were obtained from several of these patients before P. aeruginosa colonization or infection of these individuals was detected, which provided an opportunity to evaluate their responsiveness to pseudomonal exoproducts as they acquired the organism. Exotoxin A was purified from culture supernatant fluids of strain PA-103, and the two proteases were purified from an isolate of strain JR3, a highly proteolytic strain originally recovered from the sputum of a cystic fibrosis patient. Antibodies to the exotoxin A and the two proteases were detected in these sera, and sera which contained relatively high antibody levels to exotoxin A afforded mice complete protection against lethal challenges with this substance. Statistical analyses showed that patients infected with P. aeruginosa had consistently higher antibody levels (P less than 0.005) to the exoproducts than patients who were colonized with this organism. Also, patients colonized with P. aeruginosa possessed significantly higher antibody levels (P less than 0.003) to these three exoproducts than uninfected, hospitalized patients. Parke-Davis type 1 was the strain most commonly isolated from these patients (46%), but colonization or infection due to this organism usually resulted in the production of low levels of antibody to Pseudomonas exoproducts. However, infections with Parke-David type 7 organisms were always associated with intermediate- and high-responder sera to exotoxin A. These results indicated that potentially toxic products were elaborated during the course of cancer-related colonization and infection with P. aeruginosa.

ADP Ribose Transferases↗

PubMed1982

Detection of antibodies to streptococcal mucopeptide in patients with rheumatic disorders and normal controls.

Bacterial mucopeptide is an integral part of bacterial cell walls and is therefore ubiquitous in our environment. An enhanced degree of humoral immunity has ben detected not only in patients with acute rheumatic fever (ARF), with a known recent response to streptococci, but also in patients with adult and juvenile rheumatoid arthritis (RA and JRA). Our studies confirmed this association with ARF and JRA using a precipitin system as well as a radioimmunoassay to detect IgG anti-mucopeptide antibodies. In those with adult RA, either IgM or IgA rheumatoid factors or IgM or IgA antibodies specific for mucopeptide were responsible for the increased incidence of precipitins to mucopeptide in the RA patients detected in this and other studies. No differences in the specificities of the anti-mucopeptide antibodies were noted between the various patient populations as there were no lines of partial identity or nonidentity when examined by Ouchterlony double diffusion analyses. Additionally, no differences of anti-mucopeptide antibody were observed when the sera from these same patient populations were examined employing inhibition studies utilizing N-acetylglucosamine and rhamnose.

Antibodies, Bacterial↗

PubMed1981

Role of fibronectin in the prevention of adherence of Pseudomonas aeruginosa to buccal cells.

Recent evidence suggests that colonization of the upper respiratory tract by gram-negative bacilli is mediated by adherence to regional epithelial cells. Buccal epithelial cells were obtained for study from 12 seriously ill patients, all of whom were colonized with Pseudomonas aeruginosa. In comparison to cells from uncolonized controls, cells obtained from these patients attached significantly more P. aeruginosa organisms during incubation in vitro. Although the sialic acid content of colonized patients' cells was less than that of controls' cells, removal of sialic acid from normal cells with neuraminidase did not increase bacillary adherence. Trypsinization of normal cells increased bacillary adherence and significantly reduced the amount of fibronectin on the cell surface. Both trypsinized normal cells and cells recovered from seriously ill colonized patients attached large numbers of P. aeruginosa organisms in vitro and demonstrated decreased fibronectin on the cell surface by immunofluorescent staining. These findings suggest that the host alteration associated with increased susceptibility to adherence by P. aeruginosa is the loss of fibronectin from the cell surface.

Adhesiveness↗